We are using biotinylated probes for detection of target sequences.
Currently, we have started using them for detection of clones via
colony-lifts and have been experiencing background problems with the
colonies. Could this be due to cell-related biotin attracting the avidin-
alk.phos. conjugate? Is there a way to get around this problem besides
using dig-labelled probes and detection with anti-dig-alk.phos.? I've
noticed that Schleicher & Schuell's colony lift procedure with biotin-
labelled probes incorporates a step where they hand swab the lifts after
they're lysed and baked, and then incubate them in proteinase K solution.
Just curious if others have experience such background problems, or not.
Regards, Peter
*************************************************************************
* Peter M. Muriana, Ph.D. Phone = (317)-494-8284 *
* Dept. of Food Science FAX = (317)-494-7953 *
* Purdue University E-mail = muriana at aclcb.purdue.edu *
* W. Lafayette, IN 47907 *
*************************************************************************
* Disclaimer: The opinions stated above are mine alone *
* and do not represent endorsement by my employer *